Throughout human pregnancy, the placenta is indispensable for embryonic development, fetal growth and tissue differentiation. The placenta also protects the fetus against diverse insults, while preserving maternal health. Placental dysfunction is commonly implicated in complications of pregnancy that challenge maternal physiology (e.g., preeclampsia) and fetal development (e.g., fetal death or fetal growth restriction) or that leads to preterm birth. Within the placenta, the trophoblast constitutes the outermost layer, which is directly bathed in maternal blood and therefore positioned to regulate maternal-fetal gas exchange, nutrient delivery, waste removal and the production of hormones, faithfully balancing fetal needs and maternal supply. Trophoblast damage, which is common in dysfunctional placentas, may interrupt the delicate maternal-fetal balance, cause clinical disease, and leave a lifelong mark on health. A fundamental challenge in perinatal medicine arises from our limited ability to diagnose placental disorders in real time and throughout pregnancy. However, the recent discovery, by ourselves and others, that (a) placental trophoblasts release distinctive micro- and nanovesicles into the maternal circulation and (b) these vesicles contain trophoblast-specific non-coding RNA cargo, created a new opportunity for assessing trophoblast health. These vesicles are actively released by trophoblasts throughout pregnancy, and thus serve as a venipuncture-accessible natural biopsy of trophoblasts, which can furnish information on trophoblast health in real time. Our established perinatal biology group at Magee- Womens Research Institute includes expertise in perinatal medicine and placental pathology, developmental and molecular biology, and bioinformatics. Inspired by these recent advances, we have partnered with an experienced group of bioengineers that includes experts from Carnegie Mellon University, MIT, and Penn State University, with unique skills in biophysics-based vesicle analytics, including microfluidics, nanomechanics, micro/nano fabrication and vesicle sorting using acoustic tweezers. Together, our new transdisciplinary group will use integrated molecular and biophysical methodologies to directly assess the use of trophoblast-derived extracellular vesicles from maternal plasma as revelatory of trophoblast health in real time and as a technique that may be employed throughout pregnancy. Our approach is comprehensive, centering on miRNAs as well as lncRNAs and circRNAs, analyzed in exosome nanovesicles, as well as microvesicles and apoptotic bodies. As each vesicle features a unique bimolecular and biophysical signature, we will deploy our machine learning- based training and testing pipeline to informatively integrate these distinct signals into an innovative diagnostic tool. Lastly, our deployment of affordable acoustic tweezers technology to sort trophoblastic vesicles will facilitate the translation of our advances into a new lab on a chip placental diagnostic technology, suitable for small blood volumes. This technology may not only denote trophoblast pathology, but has potential to identify those who may benefit from intervention and to monitor therapeutic success.